Alzheimers Disease (AD) is a devastating neurodegenerative disease that affects millions. AD patients are found to have amyloid deposits of ABeta42 in extracellular plaques and of tau in neurofibrillary tangles. Unfortunately, amyloids are exceptionally stable and widely believed to be intractable. Moreover, soluble preamyloid oligomers may be more toxic than the fibers themselves. There are currently no therapies that prevent or reverse the formation of amyloid or the dangerous oligomeric species associated with AD, but a promising weapon against these conformers is unexpectedly found in yeast. The protein remodeling factor Hsp104 is able to rapidly disassemble yeast amyloids and remodel oligomers. Surprisingly, despite its seemingly advantageous disaggregation activity, neither Hsp104 nor a clear homolog has been identified in metazoa. The purpose of this study is to evaluate and develop Hsp104 as a tool to combat neurodegeneration associated with amyloidogenesis in AD. This will be investigated by: (1) Establishing whether Hsp104 can disaggregate and remodel aberrant ABeta42 and tau conformers. Using purified components, fibers or oligomers will be incubated with Hsp104, and disassembly will be assessed using amyloid-binding dyes, sedimentation, electron microscopy, size exclusion chromatography, and oligomer-specific antibodies. Remodeled ABeta42 and tau species will be tested for ability to seed or nucleate fiber formation, and tau will be evaluated for ability to associate with microtubules to assess possible reactivation following Hsp104 remodeling. (2) Engineering Hsp104 for optimal activity against ABeta42 and tau. Directed mutations to critical substrate-contacting Hsp104 residues may enhance substrate specificity, as monitored with the techniques above. Also, ABeta42 disassembly will be coupled to degradation by using a mutant Hsp104 that associates with the bacterial protease ClpP. Hsp104 variants will be assessed for proper assembly and ATPase activity, and ability to disaggregate of nonamyloid substrates. (3) Demonstrating that ABeta42 and tau products remodeled by Hsp104, or engineered variants, are less toxic. Both ABeta42 and tau conformers will be evaluated for toxicity by applying to cultured neuroblastoma cells. At a time when age-related neurodegenerative diseases are becoming more prevalent (and are poised to become a substantial social and economic burden), we are in desperate need of a treatment to directly eradicate amyloid and preamyloid oligomer load. Hsp104 could ultimately be employed as a gene therapy treatment for humans with AD to efficiently disassemble amyloid aggregates and neutralize toxic oligomers.